Cooperative self-assembly of discoid dimers: hierarchical formation of nanostructures with a pH switch.

Derivatives of the self-complementary 2-guanidiniocarbonyl pyrrole 5-carboxylate zwitterion (1) (previously reported by us to dimerize to 1•1 with an aggregation constant of ca. >10(10) M(-l) in DMSO) aggregate in a diverse manner depending on, e.g., variation of concentration or its protonation state. The mode of aggregation was analyzed by spectroscopic (NMR, UV) and microscopic (AFM, SEM, HIM, and TEM) methods. Aggregation of dimers of these zwitterions to higher supramolecular structures was achieved by introduction of sec-amide substituents at the 3-position, i.e., at the rearward periphery of the parent binding motif. A butyl amide substituent as in 2b enables the discoid dimers to further aggregate into one-dimensional (rod-like) stacks. Quantitative UV dilution studies showed that this aggregation is strongly cooperative following a nucleation elongation mechanism. The amide hydrogen seems to be essential for this rod-like aggregation, as neither 1 nor a corresponding tert-amide congener 2a form comparable structures. Therefore, a hydrogen bond-assisted π-π-interaction of the dimeric zwitterions is suggested to promote this aggregation mode, which is further affected by the nature of the amide substituent (e.g., steric demand), enabling the formation of bundles of strands or even two-dimensional sheets. By exploiting the zwitterionic nature of the aggregating discoid dimers, a reversible pH switch was realized: dimerization of all compounds is suppressed by protonation of the carboxylate moiety, converting the zwitterions into typical cationic amphiphiles. Accordingly, typical nanostructures like vesicles, tubes, and flat sheets are formed reversibly under acidic conditions, which reassemble into the original rod-like aggregates upon readjustment to neutral pH.

Concentration- and time-dependent effects of the synthetic estrogen, 17alpha-ethinylestradiol, on reproductive capabilities of the zebrafish, Danio rerio.

Partial or full life-cycle tests are needed to assess the potential of endocrine-disrupting compounds (EDCs) to adversely affect development and reproduction of fish. Small fish species such as zebrafish, Danio rerio, are under consideration as model organisms for appropriate test protocols. The present study examines how reproductive effects resulting from exposure of zebrafish to the synthetic estrogen 17alpha-ethinylestradiol (EE2) vary with concentration (0.05 to 10 ng EE2 L(-1), nominal), and with timing/duration of exposure (partial life-cycle, full life-cycle, and two-generation exposure). Partial life-cycle exposure of the parental (F1) generation until completion of gonad differentiation (0-75 d postfertilization, dpf) impaired juvenile growth, time to sexual maturity, adult fecundity (egg production/female/day), and adult fertilization success at 1.1 ng EE2 L(-1) and higher. Lifelong exposure of the F1 generation until 177 dpf resulted in lowest observed effect concentrations (LOECs) for time to sexual maturity, fecundity, and fertilization success identical to those of the developmental test (0-75 dpf), but the slope of the concentration-response curve was steeper. Reproduction of zebrafish was completely inhibited at 9.3 ng EE2 L(-1), and this was essentially irreversible as a 3-mo depuration restored fertilization success to only a very low rate. Accordingly, elevated endogenous vitellogenin (VTG) synthesis and degenerative changes in gonad morphology persisted in depurated zebrafish. Full life-cycle exposure of the filial (F2) generation until 162 dpf impaired growth, delayed onset of spawning and reduced fecundity and fertilization success at 2.0 ng EE2 L(-1). In conclusion, results show that the impact of estrogenic agents on zebrafish sexual development and reproductive functions as well as the reversibility of effects, varies with exposure concentration (reversibility at < or = 1.1 ng EE2 L(-1) and irreversibility at 9.3 ng EE2 L(-1)), and between partial and full life-cycle exposure (exposure to 10 ng EE2 L(-1) during critical period exerted no permanent effect on sexual differentiation, but life-cycle exposure did).

Identification of endocrine-disrupting effects in aquatic vertebrates and invertebrates: report from the European IDEA project.

The EU-funded project IDEA aimed to evaluate (a) what parameters and endpoints allow the detection of endocrine-mediated developmental and reproductive effects of (xeno)estrogens in life cycle- and life stage-specific toxicity tests with the zebrafish Danio rerio, a small laboratory fish used in many ecotoxicity test guidelines, and (b) whether substances that act as estrogens in vertebrates may also adversely affect the development, differentiation, and reproduction of aquatic invertebrates. The invertebrate species investigated included Hydra vulgaris, Gammarus pulex, Chironomus riparius, Hyalella azteca, and Lymnaea stagnalis. The animals were exposed to the model estrogenic chemicals ethynylestradiol (EE2), bisphenol A (BPA), and octylphenol (OP), which exert their endocrine activity in vertebrates through the estrogen receptor. As endpoints, developmental and reproductive parameters at the organism level as well as molecular and cellular parameters were measured. Life cycle exposure of zebrafish to (xeno)estrogens induced a specific, partly irreversible response pattern, consisting mainly of (a) induction of vitellogenin (VTG), (b) alterations of gonad differentiation, (c) delay of first spawning, and (d) reduced fertilization success. The effects of EE2 on zebrafish were expressed at environmentally realistic concentrations, while BPA and OP became effective at concentrations higher than those usually found in the environment. The vitellogenic response was equally sensitive as the reproductive parameters in the case of EE2, but VTG was more sensitive in the case of BPA. Partial life cycle exposure of zebrafish had lasting effects on fish development and reproduction only when the fish were exposed during the stage of juvenile bisexual gonad differentiation. In (partial) life cycle and multigeneration studies with invertebrates, (xeno)estrogenic impact was assessed by a range of developmental and reproductive parameters including hatching, growth, moulting, mating behavior, and egg number. Several parameters were found to be responsive to (xeno)estrogens; however, most effects were induced only at higher, probably nonphysiological concentrations. Low-dose effects were observed in full life cycle experiments, particularly in the second generation. It remains to be established whether the estrogen-induced alterations in the invertebrate species indeed do result from disturbances of the endocrine system. The findings of the present research project support the development of appropriate testing methodologies for substances with estrogenic activity.

Development and validation of a homologous zebrafish (Danio rerio Hamilton-Buchanan) vitellogenin enzyme-linked immunosorbent assay (ELISA) and its application for studies on estrogenic chemicals.

Vitellogenin (VTG) was isolated by anion exchange chromatography from plasma of female zebrafish (Danio rerio) induced with 17alpha-ethinylestradiol (EE2). The purity of the VTG isolate was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE). Purified VTG was used to raise polyclonal antibodies in rabbits and the specificity of the antisera for VTG confirmed by Western blot analysis of plasma proteins separated by SDS-PAGE. The antibodies cross-reacted with two proteins in the plasma of female zebrafish, with molecular masses of approximately 142 and 171 kDa. No cross-reactivity was observed with any other plasma proteins. A competitive enzyme-linked immunosorbent assay (ELISA) was developed using the polyclonal zebrafish VTG (z-VTG) antibodies and purified z-VTG as ligand and standard, respectively. The z-VTG ELISA was sensitive with a detection limit of between 2.0 and 3.0 ng purified VTG/ml, and a working range between 3 and 500 ng/ml (30-85% binding). The ELISA demonstrated precision, with inter- and intra-assay variations of 7.5+/-2.7 and 4.9+/-1.4%, respectively. Plasma from adult zebrafish and whole body homogenates from juvenile zebrafish diluted parallel with the z-VTG standard in the ELISA, validating the assay for quantifying z-VTG in both of these tissues. Exposure of adult male zebrafish to EE2 via water induced a concentration-dependent induction of VTG with a lowest observed effect concentration (LOEC) < or =1.67 ng EE2/l (for a 21-day exposure). The homologous z-VTG ELISA provides a valuable tool for the study of environmental estrogens in zebrafish.

Disengaging attention: on the locus of the cue-duration effect in partial report.

Increasing cue duration impairs performance in bar-probe partial report when cues are presented peripherally, but not centrally (P. Dixon, R. Gordon, A. Leung, & V. Di Lollo, 1997). Three experiments examined whether this cue-duration effect reflects processes of exogenous attention. The effect of cue duration on partial report performance with peripheral, but not central, cues was replicated (Experiment 1). Further experiments manipulated the degree that exogenous versus endogenous modes of selection were favored and found that the cue-duration effect for peripheral cues was reduced (a) when blocks contained a high proportion of central cues (Experiment 2) and (b) when the color of the cue indicated the location of the target (Experiment 3). These findings challenge the view that the cue-duration effect is restricted to exogenous attention and are discussed in terms of the process of disengaging attention from the cue to reallocate attention to the target representation.

An assessment of air toxics in Minnesota.

We used monitoring and modeling to assess the concentrations of air toxics in the state of Minnesota. Model-predicted concentrations for 148 hazardous air pollutants were from the U.S. Environmental Protection Agency Cumulative Exposure Project (1990 data). Monitoring data consisted of samples of volatile organic compounds, carbonyls, and particulate matter [Less than and equal to] 10 microm in aerodynamic diameter collected at 25 sites throughout the state for varying periods of time (up to 8 years; 1991-1998). Ten pollutants exceeded health benchmark values at one or more sites by modeling, monitoring, or both (including acrolein, arsenic, benzene, 1,3-butadiene, carbon tetrachloride, chromium, chloroform, ethylene dibromide, formaldehyde, and nickel). Polycyclic organic matter also exceeded the benzo[a]pyrene health benchmark value assumed to represent this class of pollutants. The highest modeled and monitored concentrations of most pollutants were near the center of the Minneapolis-St. Paul metropolitan area; however, many smaller cities throughout the state also had elevated concentrations. Where direct comparisons were possible, monitored values often tended to exceed model estimates. Upper-bound excess lifetime inhalation cancer risks were estimated to range from 2.7 [times] 10(-5) to 140. 9 [times] 10(-5) (modeling) and 4.7 [times] 10(-5) to 11.0 [times] 10(-5) (using a smaller set of monitored carcinogens). Screening noncancer hazard indices summed over all end points ranged from 0.2 to 58.1 (modeling) and 0.6 to 2.0 (with a smaller set of monitored pollutants). For common sets of pollutants, the concentrations, cancer risks, and noncancer hazard indices were comparable between model-based estimates and monitored values. The inhalation cancer risk was apportioned to mobile sources (54%), area sources (22%), point sources (12%), and background (12%). This study provides evidence that air toxics are a public health concern in Minnesota.

The influence of fasting on blood and plasma composition of herring gulls (Larus argentatus).

Populations of scavenging seabird species in the North Sea may fluctuate with an artificial food source: the availability of fishery waste. To document this impact, it is necessary to assess the birds' nutritional status during periods with decreased fishing activity. Reference data for this purpose was collected from 22 herring gulls investigated during laboratory fasting. After 6 d of food deprivation and body mass losses exceeding 15%, the first birds entered starvation phase 3. Comparatively, this is a rather weak fasting capacity. Plasma levels of total protein and thyroid hormones decreased and beta-hydroxybutyrate increased with fasting duration. The leucocyte proportions were shifted from lymphocytes to heterophils. After 3 d of refeeding, most of the fasting changes were reversed. Plasma enzyme activities increased and hematocrit, hemoglobin, and erythrocyte numbers decreased in both fasting and control birds, most likely as a result of experimental stress and repeated blood sampling. Glucose, cholesterol, monocytes, basophils, and glycosylated hemoglobin remained fairly constant. Triglycerides, free fatty acids, uric acid, and urea varied significantly, but changes were not as clearly a result of fasting. Therefore, total protein, beta-hydroxybutyrate, triiodothyronine, thyroxine, and lymphocyte and heterophil percentages may be the most reliable indicators of the nutritional status and the condition of free-living herring gulls.

Extramedullary facial plasmacytomas with anaplastic features: a diagnostic dilemma with implications for treatment.

Extramedullary plasmacytomas (EMPs) are rare plasma-cell tumors of the soft tissue that occur predominantly in the nasal sinuses and oropharynx. Subcutaneous and cutaneous plasmacytomas of the face are distinctly unusual. The authors report a case of rapidly expanding EMP involving the lip and contralateral nasolabial fold of a native Alaskan man with a 25-year history of recurring solitary bone plasmacytomas (SBP). An incisional biopsy revealed sheets of monotypic plasmablasts with anaplastic features. The pathologic and clinical findings were most consistent with a Richter transformation from a low-grade to a high-grade malignancy, or anaplastic myeloma (AM). With combined chemotherapy and radiation therapy, he achieved a complete response. The clinical and laboratory features of this most unusual plasma-cell dyscrasia are reviewed with an emphasis placed on diagnosis and treatment.

Role of cortisol in the ACTH-induced suppression of testicular steroidogenesis in guinea pigs.

In order to investigate the role of cortisol in the regulation of testicular function, adult male guinea pigs were challenged with ACTH (20 IU), cortisol (8 or 16 mumol), or with ACTH plus dexamethasone (DEX, 2 mumol). The amounts of cortisol, testosterone, progesterone, and androstenedione present in the plasma or secreted by incubated adrenals or testes were determined by radioimmunoassay. The plasma concentrations of LH were determined using a radioimmunoassay for rat LH. ACTH treatment elevated cortisol plasma concentrations to 999% of control values, whereas it reduced testosterone plasma levels to 43% of control values. ACTH treatment did not affect LH plasma levels. A significant negative correlation was found in ACTH-treated animals, when the cortisol and testosterone plasma concentrations in serially taken blood samples (30-240 min after treatment) were compared (rs = -0.90 and rs = -0.99, P < 0.05). In addition to cortisol, ACTH raised progesterone and androstenedione plasma concentrations. If animals were treated with 2 mumol DEX + ACTH, the plasma levels of cortisol and androstenedione but not of progesterone, testosterone or LH were changed. ACTH stimulated the in vitro secretion of cortisol, progesterone and androstenedione by the adrenals but reduced the in vitro release of androstenedione and testosterone by the testes. In summary, treatment of guinea pigs resulted in elevated cortisol and in reduced testosterone plasma concentrations. The mechanism of the cortisol-induced inhibition of testicular function was independent of the LH plasma concentrations. The in vitro experiments indicate that cortisol directly interacts with the Leydig cells, presumably by inhibiting the activity of the testicular 17 alpha-hydroxylase and/or C17,20-lyase. Taking into account the results of comparable investigations in the rat, the inhibition of the testicular 17 alpha-hydroxylase and/or C17,20-lyase takes place if the intracellular cortisol exceeds the capacity of the 11 beta-hydroxysteroid dehydrogenase to inactivate it.

Clean-up and re-use of kieselguhr (Extrelut) for liquid-liquid extraction of urinary cortisol.

Cortisol was isolated from human urine using kieselguhr (Extrelut)-filled columns. After use, Extrelut was cleaned-up once with distilled water and twice with ethanol. Before re-use, the cleaned-up kieselguhr was dried for 24 h by a warm air stream. The comparison of cortisol recovery from human urine and HPLC chromatograms of urinary extracts show that Extrelut can be repeatedly used for liquid-liquid extraction of urinary cortisol.

The use of salivary cortisol measurements for the non-invasive assessment of adrenal cortical function in guinea pigs.

For collection of saliva, cotton buds (Q-tips) were inserted into the guinea pig's cheek pouch, parallel with the cheek teeth. The recovery of saliva from the buds increased with the volume applied: whereas the recovery was 63% when 20 microliters were applied, it increased to 85% when 120 microliters were applied. The recovery of cortisol was closely related to the volume of saliva obtained by centrifugation (r = 0.99, p < 0.001). The mean value of salivary cortisol concentrations in untreated animals was 6.6 ng/ml, with relatively large variations across minutes and days within and between animals. Salivary cortisol was significantly increased if animals were singly caged, either in their familiar housing room or in an unfamiliar empty test room. In comparison to these changes, a much more pronounced increase of salivary cortisol occurred after the intramuscular (i.m.) administration of 20 IU ACTH: while the pretreatment value was 2.2 ng/ml, cortisol concentrations increased to 47 ng/ml (1 h), 72 ng/ml (2 h), 137 ng/ml (3 h) and 170 ng/ml (4 h), respectively. Similarly, i.m. administration of 2 IU insulin resulted in a significant increase of salivary cortisol (2 h: 37 ng/ml, 3 h: 24 ng/ml, 4 h: 25 ng/ml). The present study shows that the cortisol concentrations in the saliva of guinea pigs can be used as an index of adrenal cortical function in preference to the more commonly measured concentrations in the plasma. The advantages of the saliva method are: Firstly, cortisol values reflect the biologically active, unbound fraction in the plasma and are thus less affected by concentration changes of the corticosteroid-binding globulin. Secondly, saliva is easy to collect and the collection method is non-invasive; thus, no handling- or stress-induced changes of the adrenal gland occur. Thirdly, the ease of collection facilitates investigations which require frequent sampling.

Saliva cortisol and testosterone in the guinea pig: measures for the endocrine function of adrenals and testes?

For collection of saliva, cotton buds (Q-tips) were inserted into the guinea pig's cheek pouch, parallel with the cheek teeth. Within 5 min, sufficient fluid was collected for cortisol and testosterone measurements. In saline-treated animals, saliva cortisol and testosterone were about 15 ng/mL and 1.5 ng/mL, corresponding to plasma levels of 52 ng/mL and 5.9 ng/mL. Within 2-4 h after administration of 20 IU ACTH, saliva and plasma cortisol concentrations were strikingly elevated: saliva: 125 ng/mL (2 h), 157 ng/mL (4 h); plasma: 458 ng/mL (2 h), 736 ng/mL (4 h). This treatment did not influence testosterone in saliva, but reduced it in plasma (2.4 ng/mL (4 h)). In animals receiving 100 IU HCG, saliva testosterone remained unchanged, whereas its plasma levels were markedly raised (9.6 ng/mL (2 h), 12.5 ng/mL (4 h)). These results show that saliva cortisol offers promise as a noninvasive method of monitoring changes in guinea pig adrenocortical function. Saliva testosterone, on the other hand, does not correlate with plasma values; hence it cannot be used to assess testicular function in the guinea pig.

Self-care of adults with non-insulin-dependent diabetes mellitus: influence of family and friends.

This study examined the relationships among the source of support, universal self-care, and health-deviation self-care behaviors in adults who control their blood glucose with oral agents. A descriptive correlational design was used. Respondents completed a total of 51 items on questionnaires. Significant differences were found between the group with family-plus-friend support and the group without support in relation to universal self-care and health-deviation self-care. A significant difference was also found between the group with family-plus-diabetes-group support and the group without support in relation to health-deviation self-care. Subjects who received support from friends in addition to family support reported higher universal and health-deviation self-care behaviors than those without support. Support systems explained 23% of variance in universal self-care and 17% of variance in health-deviation self-care.

Measurement of salivary cortisol in guinea pigs.

For collection of saliva, cotton buds (Q-tips) were inserted into the guinea pig's cheek pouch, parallel with the cheek teeth. Within 5 minutes, sufficient fluid was collected for salivary cortisol measurements. Salivary cortisol was about 7 ng/ml after intramuscular (i.m.) injection of 0.2 ml saline. Compared with saline treatment, it was drastically increased after i.m. injection of ACTH or cortisol. Taking into account the close relationship between the amounts of cortisol in saliva and plasma in cortisol- or ACTH-treated animals, we conclude that measuring saliva cortisol offers promise as a noninvasive method to monitor the changes of adrenocortical function in guinea pigs.

Dissociation of plasma and urinary steroid values after application of stressors, insulin, vasopressin, ACTH, or dexamethasone in the Mongolian gerbil.

The amounts of cortisol and testosterone in the plasma or urine of Mongolian gerbils exposed to stress factors or treated subcutaneously with insulin (2 IU), vasopressin (1 IU), ACTH (6 IU) or dexamethasone (50 micrograms) were determined. Increased plasma cortisol was observed in animals stressed by ether anesthesia or immobilisation (1-4 hours), or treated with insulin, vasopressin or ACTH. Cortisol levels were reduced after dexamethasone administration. Plasma testosterone was elevated in animals stressed by ether anesthesia or handling plus seizure; no other treatment altered testosterone levels. An augmented cortisol excretion, which lasted one day, occurred in gerbils immobilised for one as well as for four hours. A much more prolonged stimulation of cortisol excretion, lasting three days, was seen in animals receiving ACTH or dexamethasone plus ACTH. Testosterone excretion was stimulated by ACTH and dexamethasone plus ACTH; it was not influenced by any other treatment. The present study shows that analysis of circulating steroid levels is the only reliable approach to assess the secretory activity of Mongolian gerbil adrenals or testes. In some experimental conditions (e.g. after stressor application or ACTH treatment) cortisol excretion may be used as an index of adrenal secretory function. In contrast, the striking differences between cortisol values present in plasma and urine of peptide-or dexamethasone-treated gerbils indicate that urinary cortisol does not reflect short-term changes of adrenal function. Similarly, the striking differences of testosterone values in plasma and urine indicate that urinary testosterone monitoring cannot be used to determine the secretory activity of gerbil testes.